Gene chip for identifying capripoxvirus virus and detection method of same

A gene chip, sheep pox virus technology, applied in the field of biochips, to achieve the effect of strong and stable hybridization signal, prevention of proliferation, and good specificity

Active Publication Date: 2012-03-14
重庆海关技术中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In India, people taking care of sick animals have developed red papules on t

Method used

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  • Gene chip for identifying capripoxvirus virus and detection method of same
  • Gene chip for identifying capripoxvirus virus and detection method of same
  • Gene chip for identifying capripoxvirus virus and detection method of same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] Example 1, the present invention is based on the cell proliferation of goatpoxvirus, sheeppoxvirus and bovine pimple skin disease virus of the genus Capipoxvirus, and the extraction of viral genome DNA. The ORF132 genes of goat pox virus, sheep pox virus and bovine pimple skin disease virus were cloned, sequenced and analyzed, and 8 oligonucleotide probes were designed according to the sequencing results, which can be specific for the PCR products of the corresponding viral genome DNA In combination, when performing genome amplification by PCR, the 5' end of the downstream primer is labeled with Cy3 fluorescent dye, and the probe can hybridize with this product at 56°C. The hybridization result is judged according to the position and intensity of the fluorescent signal, so as to identify goat pox virus, sheep pox virus and bovine pimple skin disease virus. PCR primers SEQ ID NO11, SEQ ID NO12 and SEQ ID NO13 were designed using Primer Premier 5.0 biological software acc...

Embodiment 2

[0081] Example 2, see figure 1 , the gene chip used for the identification of sheep poxvirus virus, adopts the micro-spotting technology of the gene chip, immobilizes the probes of the samples to be tested and various quality control probes on the chemically modified substrate, forming an 8-row×6-column array microarray. The distribution of probes on the microarray from top to bottom is as follows: spotting position quality control partition P1: 1 row × 6 points, hybridization positive quality control partition P2: 1 row × 6 points, hybridization negative quality control partition N: 1 row × 6 points, goat pox virus detection probe partition 1: 1 row × 3 points, goat pox virus detection probe partition 2: 1 row × 3 points, sheep pox virus detection probe partition 3: 1 row × 3 points, sheep pox virus Virus detection probe division 4: 1 line × 3 points, bovine pimple skin disease virus vaccine strain detection probe division 5: 1 line × 3 points, bovine pimple skin disease vir...

Embodiment 3

[0084] Embodiment 3, reagent preparation

[0085] 1) Chip washing solution

[0086] According to needs and actual conditions, prepare washing solution I and washing solution II according to the following proportions.

[0087] Washing solution Ⅰ: the final concentration of SSC is 2×, and the final concentration of SDS is 0.2%. For example, 500 mL of washing solution Ⅰ=440 mL of distilled water+50mL of 20×SSC+10mL of 10%SDS, or prepare in proportion as required.

[0088] Washing solution Ⅱ: The final concentration of SSC is 0.2×. For example, 500 mL of washing solution II=495 mL of distilled water+5 mL of 20×SSC, or prepare in proportion as required.

[0089] If 10% SDS produces a white flocculent precipitate, please dissolve it in a water bath at 42°C and mix well to prepare a lotion.

[0090] 2) Absolute ethanol.

[0091] 3) Ice-water mixture.

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Abstract

The invention discloses relates to a capripoxvirus virus, and provides a chip detecting device for goat pox virus, sheep pox virus and cow lumpy skin disease virus. In the invention, a PCR (Polymerase Chain Reaction) primer is designed through analyzing a genome sequence of a standard virus strain; a specific probe is designed through analyzing the clone and sequencing of a target gene; and the goat pox virus, the sheep pox virus and the cow lumpy skin disease virus of capripoxvirus can be identified at the same time. The invention aims at providing a method for detecting the goat pox virus, the sheep pox virus and the cow lumpy skin disease virus of capripoxvirus by using a microarray array which has the characteristics of high sensitivity, high specificity, time saving and labor saving and is easy to observe a result.

Description

technical field [0001] The invention belongs to the field of biological chips. Specifically, the present invention relates to a chip detection device for capipoxvirus (including goat poxvirus, sheep poxvirus and bovine pimple skin disease virus). Background technique [0002] Cappypox virus can cause extensive nodules and edema on the skin and organ surfaces of goats, sheep and cattle, and the milk production of sick animals will decrease sharply, and the quality of fur will greatly decrease, causing huge economic losses. Goat pox, sheep pox caused by sheep pox virus, also known as sheep "smallpox", is an acute, febrile, contact infectious disease of sheep. Sheep pox and goat pox are listed as a first-class infectious disease in my country, which has a great impact on animal husbandry. According to the difference in virulence of different strains, the lethality rate of susceptible flocks can range from 10% to 58% or from 75% to 100%. The lethality rate of lambs can reach ...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C40B40/06C12R1/93
Inventor 李应国王国民丁森王昱聂福平肖进文杨俊
Owner 重庆海关技术中心
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